Peroxisomal deficiencies are associated with altered activity of endothelial NOS in human fibroblasts.

As shown recently, in human skin fibroblasts both a constitutively expressed and the inducible nitric oxide synthase (NOS) isoform are present. To identify the NOS isoforms expressed under standard conditions in healthy human skin fibroblasts and fibroblasts with peroxisomal deficiencies (cell lines from patients suffering from X-chromosome linked Adrenoleukodystrophy (X-ALD) and the Zellweger Syndrome), we cultivated the cells in Dulbecco's modified Eagle's medium without inflammatory mediators. Our experiments clearly showed that human fibroblasts with and without peroxisomal deficiencies only contain the constitutively expressed endothelial nitric oxide synthase (eNOS) isoform and that the eNOS is tyrosine-phosphorylated. The inducible isoform (iNOS) could not be detected under standard conditions. Healthy human skin fibroblasts show a higher specific NOS activity than X-ALD and Zellweger cells (2.25 to 1.68 and 1.17 pmol L-citrulline/min/mg total cellular protein), although the content of eNOS protein does not differ significantly in these cell lines. However the tyrosine-phosphorylated portion of eNOS is significantly lower in X-ALD and Zellweger cells.